Centrifugation Application Notes

When to Use the Cell Culture Flask Adapters Cell Culture Flask Adapters support cell culture flasks during centrifugation. Centrifugation is used in the cell culturing process (seeding, passaging, or harvesting) and in studies of cellular activities (organelles, proteins, antibody production, etc.) and cellular products. During various phases (cell cycle, adhesion, motility, signal transduction, etc.) of the cellular study, centrifugation is used to separate or concentrate cells from culture media and/or to collect extracellular products. Traditionally, the cells and media are transferred from a cell culture flask to a container (typically a 15 mL or 50 mL conical tube). After centrifugation, a cell pellet is either resuspended and transferred back to the cell culture flask for continued growth, or it is extracted for storage or downstream analysis. These transfer steps can be eliminated when culturing and centrifugation are carried out in the same flask using the Cell Culture Flask Adapters. The streamlined process is applicable for various types of cell cultures (adherent, suspension, hybridoma, and primary cells; see Fig. 2).

CENTRIFUGATION STEPS IN THE CELL CULTURING PROCESS Each transfer step in red italics eliminated with the Cell Culture Flask Adapter.

Thawing

Start with primary cell line from blood, tissues…

Thaw frozen established cell line

Starting

Seeding

Preliminary Separation

Resuspend cells in growing media, grow cells in Cell Culture Flask

ADHERENT CELLS

SUSPENSION CELLS

Expansion

Multiple Passaging 1. Transfer from Flask to Conical Tube 2. Centrifuge to Pellet 3. Recover Cells or Supernatant 4. Replenish Media and Continue Growth

Trypsin Treatment/Multiple Passaging 1. Recover Supernatant 2. Add Trypsin to Dislodge Cells 3. Transfer from Flask to Conical Tube 4. Rinse/Wash Cells 5. Centrifuge to Pellet 6. Recover Cells 7. Replenish Media and Continue Growth

Multiple Passaging

Harvesting Cell

Harvesting/Adjusting Cell Concentration 1. Transfer from Flask to Conical Tube 2. Centrifuge to Pellet 3. Recover Cell/Discard Supernatant 4. Resuspend in Media/Adjust Cell Concentration 5. Transfer to Labware

Cell Storage

Downstream Assay

Figure 2. Centrifugation steps in the cell culturing process.

2